Hasche K D, Schaeg W, Blobel H, Brückler J
Zentralbl Bakteriol Orig A. 1977 Jul;238(3):300-9.
380 (80%) of 475 Staphylococcus aureus cultures isolated from humans, cattle and dogs were proteolytically active either on casein or gelatin or both (table 1). Protease-activity could also be demonstrated in experimental body-cavities of rabbits (fig. 1). The enzyme-activity was estimated with azocasein. Protease from S. aureus, M 135 precipitated from the culture supernatant with ammonium sulfate at 65% saturation (table 2). It was purified by 2 filtrations on Ultrogel AcA 44 (fig. 2,3) and subsequent isoelectric focusing between pH 3.5-7.0 (fig. 4). The purified protease yielded only 1 line in the SDS-polyacrylamidegel-electrophoresis, in the gelatin-polyacrylamidegel-electrophoresis and in the double immuno-diffusion test (fig. 5). Its isoelectric point was at pH 4.6, and its highest proteolytic activity between pH 7.5-8.3. The molecular weight was estimated by SDS-polyacrylamidegel-electrophoresis to be near 29.000. The protease-activity was completely inhibited in the presence of EDTA, partially inhibited by Cu2+ and Zn2+ and increased by Mn2+ (table 3).
从人、牛和狗身上分离出的475株金黄色葡萄球菌培养物中,有380株(80%)对酪蛋白或明胶或两者都具有蛋白水解活性(表1)。在兔的实验体腔中也可证明蛋白酶活性(图1)。用偶氮酪蛋白估计酶活性。来自金黄色葡萄球菌的蛋白酶,M 135在65%饱和度的硫酸铵作用下从培养上清液中沉淀出来(表2)。通过在Ultrogel AcA 44上进行2次过滤(图2、3)以及随后在pH 3.5 - 7.0之间进行等电聚焦(图4)进行纯化。纯化后的蛋白酶在SDS - 聚丙烯酰胺凝胶电泳、明胶 - 聚丙烯酰胺凝胶电泳和双向免疫扩散试验中仅产生1条带(图5)。其等电点为pH 4.6,在pH 7.5 - 8.3之间具有最高的蛋白水解活性。通过SDS - 聚丙烯酰胺凝胶电泳估计分子量接近29000。在EDTA存在下蛋白酶活性完全被抑制,被Cu2 +和Zn2 +部分抑制,被Mn2 +增强(表3)。